Assays for Gene Transfer

Abstract

There are, of course, several methods that may be employed to monitor and quantitate successful gene transfer. One can assay for the presence of transferred DNA by a Southern blot of genomic DNA or, if the transferred DNA has some unique sequence feature, by the polymerase chain reaction (PCR) carried out on genomic DNA. One may also monitor successful gene transfer by assaying for mRNA production by the exogenous DNA through Northern analysis of cellular RNA or by RNA-driven PCR. Similarly, protein production can be monitored through the application of either immune precipitation or Western techniques, as well as immunofluorescence analysis or bioassay where appropriate. In this section on assays for gene transfer, I discuss and describe Southern- and PCR-based DNA-detection techniques. I will leave RNA- and protein-detection methods for the next section.